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目的:探究野生云芝多糖水溶性新组分CVPS-B对大鼠脾细胞单核细胞趋化蛋白-1(MCP-1)基因表达的影响.方法:以β-actin为内标准物,用逆转录聚合酶链式反应(RT-PCR)检测CVPS-B分别对正常情况下以及脂多糖(LPS)诱导下大鼠脾细胞MCP-I基因表达的影响,并对RT-PCR产物进行测序,以证实其特异性.结果:(1)正常情况下大鼠脾细胞MCP-1 mRNA的表达(MCP-1/β-actin的比值)生理盐水对照组为1.4±0.3;CVPS-B三个剂量组(10、30和50mg·kg~(-1)·d~(-1),ip,连续4d)分别为:1.6±0.4、1.7±0.5和1.5±0.4,后三组与对照组无显著差异(P>0.05);(2)大鼠腹腔给药LPS(10μg·kg~(-1)可使脾细胞MCP-1 mRNA的表达增加114％.(3)CVPS-B4个剂量组(5、10、30和50mg·kg~(-1)·d~(-1),ip,连续4d)可使LPS(10μg·kg~(-1),ip)诱导的脾细胞MCP-1 mRNA的表达分别减低51％,70％,84％和99％(n=6).结论:CVPS-B可预防性抑制LPS对大鼠脾细胞MCP-1基因表达的诱导作用,且呈剂量依赖性,但对正常情况下大鼠脾细胞MCP-1 mRNA的表达则无明显影响.
Objective: To investigate the effect of CVPS-B, a water-soluble new component of wild kudzu polysaccharide, on the expression of rat monocyte chemoattractant protein-1 (MCP-1) gene. Methods: β-actin was used as an internal standard, Reverse transcriptase polymerase chain reaction (RT-PCR) was used to detect the effect of CVPS-B on the expression of MCP-I gene in spleen cells induced by lipopolysaccharide (LPS) under normal conditions, and the RT-PCR products were sequenced. To confirm its specificity. Results: (1) Normally, the expression of MCP-1 mRNA in rat spleen cells (ratio of MCP-1/β-actin) was 1.4±0.3 in saline control group; three doses of CVPS-B were used. The groups (10, 30 and 50 mg·kg -1 ·d -1), ip, continuous for 4 days were: 1.6±0.4, 1.7±0.5 and 1.5±0.4, respectively, and there was no significant difference between the latter three groups and the control group. Differences (P>0.05); (2) LPS (10 μg·kg -1 ) increased the expression of MCP-1 mRNA in splenocytes by 114%. (3) CVPS-B4 doses (5) 10, 30, and 50 mg·kg -1 ·d -1 ip for 4d consecutively induced LPS (10 μg·kg -1, ip) induced MCP-1 mRNA expression in splenocytes Expression decreased by 51%, 70%, 84%, and 99%, respectively (n=6). Conclusion: CVPS-B can prevent LPS-induced induction of MCP-1 gene expression in rat spleen cells in a dose-dependent manner. Sex, but no significant effect on the expression of MCP-1 mRNA in spleen cells of normal rats.